Site-specific effects of intracranial estradiol administration on recurrent insulin-induced hypoglycemia in the ovariectomized female rat

Clinical and experimental studies reveal gender differences in susceptibility to dampening effects of precedent hypoglycemia on recurrent insulin-induced hypoglycemia (RIIH). Recent studies implicate the ovarian steroid, estradiol, in the regulation of RIIH, since systemic replacement of this hormone at basal estrous cycle levels maintains glucose profiles during serial insulin dosing and prevents RIIH-associated reductions in neuronal activation in key metabolic structures in the ovariectomized female rat brain. The present study investigated the hypothesis that these effects are achieved, in part, by estrogenic action within the central nervous system, including glucoregulatory structures characterized by high estrogen receptor (ER) expression. Initial experiments evaluated the impact of global intracranial administration of estradiol on RIIH. Ovariectomized rats were treated by continuous infusion of graded doses of 17β-estradiol-3-benzoate (EB) or vehicle into the lateral ventricle (LV), and injected subcutaneously with 1 or 4 doses of the intermediate-release insulin, Humulin N (HN), 1 dose per day. Animals infused with 5 or 10 µg EB/day exhibited uniform glycemic responses to 1 versus 4 doses of insulin, whereas rescue from hypoglycemia was delayed during repetitive HN injection of rats infused with either vehicle or 1 µg EB/day. Recovery from both single and multiple bouts of hypoglycemia was more rapid in rats infused with the higher EB doses, compared to other groups. Mapping of ERα immunoreactivity in animals treated by LV infusion of EB revealed variable nuclear staining in ER-expressing metabolic loci typified by estrogen-dependent sustenance of neuronal reactivity to hypoglycemia, with highest levels of ERα immunoreactivity observed in the arcuate (ARH) and ventromedial (VMH) hypothalamic nuclei, and moderate labeling of the caudal hindbrain dorsal vagal complex. EB delivery to the caudal hindbrain via the caudal fourth ventricle resulted in dose-dependent effects on RIIH, since glycemic profiles were either unchanged or diminished relative to acute NH-induced hypoglycemia, in high versus low EB-treated animals, respectively. Bilateral administration of 1.0 µg EB into the ARH or VMH elicited disparate effects on acute and chronic HN-induced hypoglycemia. Intra-VMH EB delayed recovery from both acute and chronic hypoglycemia, compared to non-estradiol-treated controls. In contrast, neither that dose nor a 10-fold lower dosage of EB delivered to the ARH modified acute HN-induced hypoglycemia, but RIIH was either attenuated or enhanced, respectively, in animals treated by intra-ARH delivery of 1.0 versus 0.1 µg EB, respectively. These results suggest that whole brain exposure to elevated estradiol may promote outflow that truncates hypoglycemia and maintains glucose profiles during RIIH, whereas actions of relatively low hormone levels on the central nervous system may result in adaptive adjustments that result in lower blood glucose levels during recurring versus acute hypoglycemia. The data also imply that, at a given concentration, estrogens may exert site-specific effects on acute and chronic HN-induced hypoglycemia. Further research is needed to identify the cellular substrates and physiological mechanisms that mediate caudal hindbrain-, ARH-, and VMH-specific actions of estradiol on acute and chronic hypoglycemia.